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The FilmArray® Gastrointestinal (GI) Panel is a modern, sensitive, and comprehensive stool testing technique for identifying common gastrointestinal pathogens, including viruses, bacteria, and parasites. Its increasing demand is due to ease of operation and automation. Pathogens, particularly viruses, undergo constant genetic evolution. For instance, human astrovirus (HAstV), which causes gastroenteritis in children, the elderly, and immune-compromised individuals, can be identified by the GI Panel. HAstV has evolved into several clades, including the classic (HAstV1-8), novel Melbourne (MLB1-3), and Virginia (VA1-5) clades. This study investigated whether the GI Panel accurately detects all HAstV clades. A total of 12 stool and three sewage water (SW) samples were selected post-confirmation of distinct HAstV strains using conventional RT-PCR and sequence-based genotyping for reassessment by the GI Panel. The GI Panel accurately detected the classic HAstV in stool and SW samples. However, our results confirm the GI Panel's inability to detect the novel MLB (MLB1-3) and VA (VA2) clades in fecal samples, raising the possibility of false-negative results in HAstV testing. Although the GI Panel is useful for identifying a variety of gastrointestinal pathogens in stool and SW samples in a single test, our findings highlight the need to exercise caution when interpreting HAstV results from the GI Panel.
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Noroviruses (NoVs) are a global concern, causing widespread outbreaks and sporadic acute gastroenteritis (AGE) cases across all age groups. Recent research has shed light on the emergence of novel recombinant strains of NoV in various countries. To delve deeper into this phenomenon, we extensively analyzed 1,175 stool samples collected from Japanese infants and children with AGE from six different prefectures in Japan over three years, from July 2018 to June 2021. Our investigation aimed to determine the prevalence and genetic characteristics of NoV associated with sporadic AGE while exploring the possibility of detecting NoV recombination events. Among the analyzed samples, we identified 355 cases positive for NoV, 11 cases attributed to GI genotypes, and 344 associated with GII genotypes. Notably, we discovered four distinct GI genotypes (GI.2, GI.3, GI.4, and GI.6) and seven diverse GII genotypes (GII.2, GII.3, GII.4, GII.6, GII.7, GII.14, and GII.17). The predominant genotypes were GII.4 (56.4%; 194 out of 344), followed by GII.2 and GII.3. Through dual genotyping based on sequencing of the ORF1/ORF2 junction region, we identified a total of 14 different RdRp/capsid genotypes. Of particular interest were the prevalent recombinant genotypes GII.4[P31] and GII.2[P16]. Notably, our study revealed a decrease in the number of children infected with NoV during and after the COVID-19 pandemic. These findings underscore the importance of continuous NoV surveillance efforts.
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Infecções por Caliciviridae , Variação Genética , Norovirus , Criança , Pré-Escolar , Humanos , Lactente , Infecções por Caliciviridae/epidemiologia , Infecções por Caliciviridae/virologia , COVID-19 , Fezes/virologia , Genótipo , Japão/epidemiologia , Norovirus/classificação , Norovirus/genética , Filogenia , Prevalência , Adolescente , Proteínas do Capsídeo/genéticaRESUMO
Mycobacterium tuberculosis (Mtb) infection remains a major health problem worldwide. Although the Bacillus Calmette-Guérin (BCG) vaccine is the most widely used vaccination for preventing tuberculosis (TB), its efficacy is limited. We previously developed a new recombinant BCG (rBCG)-based vaccine encoding the Ag85B protein of M. kansasii (Mkan85B), termed rBCG-Mkan85B, and its administration is followed by boosting with plasmid DNA expressing the Ag85B gene (DNA-Mkan85B). Previously, we identified MHC-I (H2-Kd)-restricted epitopes that highly cross-react with those of Mtb in BALB/c (H2d) and CB6F1 (H2b/d) mice. We also reported that the rBCG-Mkan85B/DNA-Mkan85B prime-boost vaccination protocol protected CB6F1 mice against M. kansasii infection. In this study, to investigate the protective effect of our novel rBCG against Mtb infection, CB6F1 mice were either left unimmunized or immunized with the BCG, rBCG-Mkan85B, or rBCG-Mkan85B/DNA-Mkan85B vaccine for 10 weeks prior to inhalation exposure to the virulent Mtb Erdman strain for another 6 weeks. Compared with the BCG and rBCG-Mkan85B vaccinations, the rBCG-Mkan85B/DNA-Mkan85B prime-boost vaccination protocol significantly reduced the numbers of pulmonary colony-forming units (CFUs). Moreover, the rBCG-Mkan85B/DNA-Mkan85B prime-boost vaccination induced antigen-specific polyfunctional CD4+ and CD8+ T cells. These results suggest that CD8+ T-cell immunity to immunodominant epitopes of Mtb is enhanced by rBCG vector-based immunization. Thus, rBCG vector-based vaccinations may overcome the limited ability of the current BCG vaccine to elicit TB immunity.
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Mycobacterium bovis , Mycobacterium tuberculosis , Tuberculose , Animais , Camundongos , Vacina BCG , Linfócitos T CD8-Positivos , Antígenos de Bactérias , Proteínas de Bactérias/metabolismo , Mycobacterium tuberculosis/metabolismo , Camundongos Endogâmicos BALB CRESUMO
Transforming growth factor-beta 1 (TGF-ß1) is a pleiotropic growth factor playing various roles in the human body including cell growth and development. More functions of TGF-ß1 have been discovered, especially its roles in viral infection. TGF-ß1 is abundant at the maternal-fetal interface during pregnancy and plays an important function in immune tolerance, an essential key factor for pregnancy success. It plays some critical roles in viral infection in pregnancy, such as its effects on the infection and replication of human cytomegalovirus in syncytiotrophoblasts. Interestingly, its role in the enhancement of Zika virus (ZIKV) infection and replication in first-trimester trophoblasts has recently been reported. The above up-to-date findings have opened one of the promising approaches to studying the mechanisms of viral infection during pregnancy with links to corresponding congenital syndromes. In this article, we review our current and recent advances in understanding the roles of TGF-ß1 in viral infection. Our discussion focuses on viral infection during pregnancy, especially in the first trimester. We highlight the mutual roles of viral infection and TGF-ß1 in specific contexts and possible functions of the Smad pathway in viral infection, with a special note on ZIKV infection. In addition, we discuss promising approaches to performing further studies on this topic.
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Infecção por Zika virus , Zika virus , Gravidez , Feminino , Humanos , Fator de Crescimento Transformador beta1/metabolismo , Zika virus/metabolismo , Primeiro Trimestre da Gravidez , Trofoblastos/metabolismoRESUMO
To control the coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the promotion of vaccination is important. However, adverse reactions following vaccination remain a concern. To investigate adverse events in the vaccinated Japanese population, we conducted a survey-based study among health care workers, including medical doctors and nurses; other medical staff; and medical university faculty, staff, and students in a single medical school and affiliated hospital in Japan. In addition, we analyzed the association of different adverse events with individual factors (e.g., age, sex) by performing network analysis. While young age and female sex are often considered risk factors for more severe adverse events, the regression models showed neither age nor sex was associated with local injection-site reactions after the second dose in this study. In contrast to local reactions, systemic adverse events were associated with young age and female sex. However, myalgia was unique in that it was not associated with younger age even though the network analysis showed that myalgia was consistently related to arthralgia and belonged to the group of systemic events after both the first and second vaccine doses. Further study is needed to ensure safe and effective vaccination to aid in controlling the COVID-19 pandemic.
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Vacina BNT162 , COVID-19 , Pessoal de Saúde , Estudantes de Medicina , Vacina BNT162/efeitos adversos , COVID-19/epidemiologia , COVID-19/prevenção & controle , Feminino , Humanos , Japão/epidemiologia , Mialgia/induzido quimicamente , Mialgia/epidemiologia , SARS-CoV-2 , Vacinação/efeitos adversosRESUMO
INTRODUCTION: Norovirus (NoV) is the most common agent causing outbreaks and sporadic cases of acute gastroenteritis among all ages, especially children under 5 years old. During the coronavirus disease 2019 (COVID-19) pandemic, NoV infection has decreased drastically in Japan due to school closures and no outbreak related to NoV infection had been reported. METHOD: In mid-September 2021, NoV outbreak occurred in kindergarten and nursery schools in Maizuru, Kyoto prefecture, Japan. Twenty-six stool samples collected from patients who were diagnosed of NoV gastroenteritis from the outbreak by an immunochromatographic (IC) kit at a pediatric outpatient clinic in Maizuru city during 3 weeks from September 13 to October 8, 2021 were examined for the presence of NoV GII by reverse transcriptase-polymerase chain reaction (RT-PCR), genome sequencing, and phylogenetic analysis. RESULT: All 26 samples were confirmed positive to NoV GII and their genotypes were identified as GII.4 Sydney[P31]. The amino acid substitutions in open reading frame1 (ORF1) and ORF2 genes were found when compared with previously detected sporadic NoV GII.4 Sydney[P31] strains isolated in Japan. The clinical characterization of infected children was described. Most of the children were mild cases and vomiting was the most frequent clinical symptom. CONCLUSION: This study reported a recent emergence of NoV GII.4 Sydney[P31] causing acute gastroenteritis outbreak in children in Japan during the COVID-19 pandemic and suggests a need for further monitoring of NoV GII.4 variants.
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COVID-19 , Infecções por Caliciviridae , Gastroenterite , Norovirus , COVID-19/epidemiologia , Infecções por Caliciviridae/epidemiologia , Criança , Pré-Escolar , Fezes , Gastroenterite/epidemiologia , Genótipo , Humanos , Japão/epidemiologia , Norovirus/genética , Pandemias , FilogeniaRESUMO
Although various perinatal outcomes in coronavirus disease 2019 (COVID-19) pregnancies have been reported, the fetal and neonatal consequences of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection remain unclear. Several reports of miscarriages and stillbirths have been recorded, but vertical transmission by SARS-CoV-2 is considered very rare, and the cause remains unknown. We report a case of a 22-year-old uncomplicated Japanese woman infected with SARS-CoV-2 during the second trimester, resulting in intrauterine fetal death due to placental insufficiency associated with COVID-19 placentitis. This report emphasizes the importance of longitudinal assessment of fetal well-being by fetal heart rate monitoring and early detection of maternal coagulation dysfunction representing SARS-CoV-2 inflammation to manage COVID-19 in pregnancy.
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Transtornos da Coagulação Sanguínea , COVID-19 , Corioamnionite , Complicações Infecciosas na Gravidez , Adulto , COVID-19/complicações , Feminino , Morte Fetal/etiologia , Frequência Cardíaca Fetal , Humanos , Recém-Nascido , Transmissão Vertical de Doenças Infecciosas , Placenta , Gravidez , Resultado da Gravidez , SARS-CoV-2 , Natimorto , Adulto JovemRESUMO
BACKGROUND: The COVID-19 vaccine is effective in preventing severe cases of COVID-19. For women, gynecological adverse events, such as menstrual irregularities and irregular bleeding, could be a concern after COVID-19 vaccination. In this study, we investigated gynecological adverse events in the vaccinated Japanese female population. METHODS: We conducted a survey-based study with health-care workers, including medical doctors and nurses, medical coworkers, and medical university faculty, staff, and students, at a single medical school and affiliated hospital in Japan. We used McNemar's test and network analysis. RESULTS: Overall, we obtained 819 responses, and 424 were from females. After the exclusion of contradictory answers, 309 surveys were finally considered appropriate for the analysis. The frequencies of abnormal bleeding were 0.6%, 1.0%, and 3.0% for the first, second, and third doses, respectively. An irregular menstrual cycle was more common than abnormal bleeding: 1.9%, 4.9%, and 6.6% for the first, second, and third doses, respectively. Network analysis revealed that abnormal bleeding and an irregular menstrual cycle were not associated with other adverse reactions. CONCLUSION: The present study showed that the effects of COVID-19 vaccination on menstruation seem limited.
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Vacinas contra COVID-19 , COVID-19 , Feminino , Humanos , Vacina BNT162 , Distúrbios Menstruais , Ciclo Menstrual , Vacinas de mRNARESUMO
To investigate the vaccination status and adverse reactions to the COVID-19 vaccine among pregnant women in Japan, we conducted an online questionnaire survey from October 5 to November 22, 2021. The number of participants in the online survey was 6576. Of the participants, 4840 (73.6%) were vaccinated twice, and 557 (8.5%) were vaccinated once. A total of 1179 (17.9%) responders had never been vaccinated against COVID-19. The most frequent adverse reaction was local pain at the injection site. The incidence of local adverse reactions was almost identical after the first and the second vaccinations, while systemic reactions, such as fever and fatigue/malaise, and adverse reactions outside the vaccination site such as headache and arthralgia, were more frequent after the second vaccination than after the first vaccination. Regarding the obstetrical complications, uterine tension and/or contraction was observed in 1.65% of the pregnant women after the first vaccination and in 2.98% after the second vaccination, and uterine pain appeared in 1.06% of the pregnant women after the second vaccination. However, serious symptoms, such as hemorrhage, decreased fetal movement, edema, increased blood pressure, and amniorrhexis, were seen in less than 1% of vaccinated women after both the first and second vaccinations. This study clarified the characteristics of vaccination, adverse reactions, and obstetrical symptoms in pregnant women in Japan who had the COVID-19 vaccine up to the second dose. As a booster vaccination is currently underway, further study is needed to improve the management of pregnant women during the current pandemic.
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Vacinas contra COVID-19 , COVID-19 , Gestantes , COVID-19/prevenção & controle , Vacinas contra COVID-19/efeitos adversos , Feminino , Humanos , Japão/epidemiologia , Dor/etiologia , Gravidez , Vacinação/efeitos adversosRESUMO
INTRODUCTION: Recently, increased frequencies of carbapenemase-producing Enterobacteriaceae have been reported worldwide. Among multiple genetic subtypes, oxacillinase (OXA)-48 ß-lactamase-producing strains have been associated with inbound infection because they have been detected predominantly in patients who traveled outside of Japan. However, a recent case report of OXA-48 ß-lactamase-producing Enterobacteriaceae suggested the latent spread of domestic infections. Due to a lack of specific inhibitors, culture-based detection of OXA-48 ß-lactamase-producing bacteria is difficult. Thus, DNA-based detection methods, including PCR, direct sequencing and loop-mediated isothermal amplification (LAMP), have been employed. Among these methods, LAMP detection is more favorable than other methods because of its technical simplicity and low cost. METHODS: We designed novel LAMP primers to detect OXA-48 ß-lactamase-producing bacteria and investigated their possible clinical applications with bacterial genome-spiked human materials (cerebrospinal fluid, blood, feces, urine, and sputum). We evaluated the specificity of the LAMP primers using 37 bacterial strains: 8 standard, 9 reference, and 20 clinical Gram-negative strains. RESULTS: Our LAMP primers detected 10 copies of the OXA-48 type ß-lactamase gene and exhibited no cross reactivity with other ß-lactamase genes. Sensitivity was not influenced in any clinical sample, in contrast to PCR detection, which was strongly inhibited by substances in fecal samples. CONCLUSIONS: These results suggest the superior performance of LAMP compared with conventional PCR for detecting the OXA-48 type ß-lactamase gene in various clinical samples.
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Técnicas de Amplificação de Ácido Nucleico , beta-Lactamases , Proteínas de Bactérias/genética , Bactérias Gram-Negativas/genética , Humanos , Japão , Técnicas de Diagnóstico Molecular , Sensibilidade e Especificidade , beta-Lactamases/genéticaRESUMO
The current COVID-19 pandemic is a global concern. The recent introduction of vaccines has provided a reason for hope, but new problems, such as vaccine hesitancy, have arisen. One of the most important of these issues is the safety of vaccines for pregnant women. In this article, we collected worldwide indications for vaccination, including women who are pregnant or who wish to become pregnant, and reports of adverse reactions to COVID-19 vaccination. The Japan Society of Obstetrics and Gynecology and the Japanese Society of Infectious Diseases in Obstetrics and Gynecology have published recommendations for the vaccination of pregnant women with a COVID-19 vaccine. The guidelines are as follows: (1) pregnant women should not be excluded from vaccination; (2) informed consent should be obtained before vaccination; (3) healthcare workers and pregnant women with complications such as diabetes, hypertension, and obesity should be vaccinated preferentially; (4) vaccination should be avoided until 12 weeks of gestation during organogenesis; (5) spouse and family members should be vaccinated actively; and (6) nursing mothers are not particularly affected. This policy has been adopted in government guidelines. Additional efforts should be made to protect pregnant women from infection and severe illness with COVID-19 by eliminating vaccine hesitancy.
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COVID-19 , Complicações Infecciosas na Gravidez , Vacinas contra COVID-19 , Feminino , Humanos , Japão , Pandemias , Gravidez , Complicações Infecciosas na Gravidez/prevenção & controle , Gestantes , SARS-CoV-2 , VacinaçãoRESUMO
Despite efforts to develop effective treatments and vaccines, Mycobacterium tuberculosis (Mtb), particularly pulmonary Mtb, continues to provide major health challenges worldwide. To improve immunization against the persistent health challenge of Mtb infection, we have studied the CD8+ T cell response to Bacillus Calmette-Guérin (BCG) and recombinant BCG (rBCG) in mice. Here, we generated CD8+ T cells with an rBCG-based vaccine encoding the Ag85B protein of M. kansasii, termed rBCG-Mkan85B, followed by boosting with plasmid DNA expressing the Ag85B gene (DNA-Mkan85B). We identified two MHC-I (H2-Kd )-restricted epitopes that induce cross-reactive responses to Mtb and other related mycobacteria in both BALB/c (H2d ) and CB6F1 (H2b/d ) mice. The H2-Kd -restricted peptide epitopes elicited polyfunctional CD8+ T cell responses that were also highly cross-reactive with those of other proteins of the Ag85 complex. Tetramer staining indicated that the two H2-Kd -restricted epitopes elicit distinct CD8+ T cell populations, a result explained by the X-ray structure of the two peptide/H2-Kd complexes. These results suggest that rBCG-Mkan85B vector-based immunization and DNA-Mkan85B boost may enhance CD8+ T cell response to Mtb, and might help to overcome the limited effectiveness of the current BCG in eliciting tuberculosis immunity.
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Aciltransferases/imunologia , Antígenos de Bactérias/imunologia , Vacina BCG/imunologia , Proteínas de Bactérias/imunologia , Linfócitos T CD8-Positivos/imunologia , Antígenos de Histocompatibilidade Classe I/metabolismo , Tuberculose/imunologia , Vacinas de DNA/imunologia , Sequência de Aminoácidos , Animais , Epitopos/imunologia , Feminino , Imunização/métodos , Imunização Secundária/métodos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Mycobacterium bovis/imunologia , Mycobacterium tuberculosis/imunologia , Tuberculose/microbiologia , Vacinação/métodosRESUMO
At the end of 2019, a new coronavirus disease, COVID-19, emerged and quickly spread around the world. Severe acute respiratory syndrome Coronavirus 2 (SARS-CoV-2), the causative virus of this disease, belongs to the ß-coronavirus family, together with SARS and middle east respiratory syndrome, and has similar biological characteristics to these viruses. For obstetricians, the susceptibility and prognoses of pregnant women and the effects of the infection on the fetus have been the focus of attention; however, at present, the seriousness of the disease in pregnant women is not apparent, and COVID-19 does not increase the rate of miscarriage, stillbirth, preterm labor or teratogenicity. Even so, carriers might transmit SARS-CoV-2 to pregnant women. Thus, we must keep in mind that all medical personnel must understand and maintain standard precautions in their clinical and laboratory practices.
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Betacoronavirus , Infecções por Coronavirus/transmissão , Transmissão Vertical de Doenças Infecciosas/prevenção & controle , Pneumonia Viral/transmissão , Complicações Infecciosas na Gravidez/virologia , COVID-19 , Feminino , Humanos , Pandemias , Gravidez , Complicações Infecciosas na Gravidez/epidemiologia , SARS-CoV-2RESUMO
From last decade of the 20th century, numerous epidemiological studies and intervention trials have attempted to prove the relationships between maternal periodontal diseases and adverse pregnancy outcomes (APO). Periodontal diseases are considered a risk factor for APO, including preterm birth, fetal growth restriction, low birthweight, pre-eclampsia and gestational diabetes. However, the efficacy of periodontal treatment during pregnancy is controversial. Two pathogenic mechanisms might explain the potential effect of periodontal diseases on pregnancy outcomes. First, periodontal bacteria originating in the gingival biofilm directly affect the feto-placental unit subsequent to bacteremia. Second, inflammatory mediators secreted by the subgingival inflammatory site are carried to the feto-placental unit, where they then cause an inflammatory response. To elucidate these mechanisms, many researchers have been investigating the use of experimental animal models and in vitro models. In the present review, we summarize the current literature on the relationship between periodontal diseases and APO from epidemiological studies, animal models studies and in vitro studies, and speculate on the possible mechanism of periodontal diseases affecting pregnancy outcomes.
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Recém-Nascido de Baixo Peso , Doenças Periodontais , Pré-Eclâmpsia , Complicações na Gravidez , Nascimento Prematuro , Animais , Feminino , Humanos , Doenças Periodontais/complicações , Doenças Periodontais/imunologia , Doenças Periodontais/microbiologia , Pré-Eclâmpsia/etiologia , Gravidez , Complicações na Gravidez/etiologia , Complicações na Gravidez/imunologia , Complicações na Gravidez/microbiologia , Nascimento Prematuro/etiologiaRESUMO
Extended-spectrum beta-lactamase (ESBL) producing bacteria spread worldwide and became major concern for antibiotic treatment. Although surveillance reports in general hospitals and long-term care facilities are increasing, their frequencies in individuals with severe motor and intellectual disabilities (SMID) are so far unknown. In this study, we examined the frequency of ESBL in stool samples collected from 146 asymptomatic SMID subjects hospitalized in a single institution. With their clinical information, we evaluated possible risk factors for ESBL colonization. From 146 fecal samples, ESBL-producing bacteria were isolated in 45 cases (31%). Drug sensitivity testing showed that 82% of the isolates were resistant to levofloxacin but were sensitive to tazobactam/piperacillin and cefmetazole. The most frequent genotype was CTX-M-9 detected in 36/45 (80%). A high degree of disability, antibiotic use within three months before sampling and post-tracheostomy were statistically significant risk factors. Tube feeding was also strongly correlated with ESBL colonization (p < 0.001) and associated with lower micro-organismic diversities. Our findings are the first to reveal a high prevalence of ESBL in the fecal samples of SMID individuals and suggest possible relationships between high degree disability, tube feeding and latest histories of antibiotic use.
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Proteínas de Escherichia coli/isolamento & purificação , Fezes/microbiologia , Deficiência Intelectual/microbiologia , Microbiota/genética , Transtornos Motores/microbiologia , beta-Lactamases/isolamento & purificação , Adolescente , Adulto , Idoso , Antibacterianos/metabolismo , Criança , Pré-Escolar , Nutrição Enteral , Infecções por Enterobacteriaceae/microbiologia , Proteínas de Escherichia coli/genética , Humanos , Pessoa de Meia-Idade , Prevalência , Fatores de Risco , Traqueostomia , beta-Lactamases/genéticaRESUMO
BACKGROUND: Delayed wound healing reduces the quality of life (QOL) of patients. Thus, understanding the mechanism of wound healing is indispensable for better management. However, the role of innate immunity in wound healing is thus far unknown. Recently the involvement of TLR3 in wound healing has been evaluated. The systemic administration of polyriboinosinic-polyribocytidylic acid (poly I:C ; a substitute for viral dsRNA and a ligand of toll-like receptor 3), enhances wound healing in vivo. The aim of this study is to improve our understanding of the link between innate immunity and human wound healing, particularly in re-epithelialization. RESULTS: The present study showed that poly I:C significantly accelerated collective HaCaT cell migration in a scratch assay. Poly I:C also increased IL-8 and bFGF production, and anti-IL-8 antibodies significantly inhibited the migration caused by poly I:C. Human recombinant IL-8 also accelerated collective HaCaT cell migration. An immunofluorescence assay and enzyme-linked immunosorbent assay (ELISA) also revealed that poly I:C decreased E-cadherin protein levels and increased vimentin protein levels, and anti-IL-8 antibody reversed this effect. In contrast, nucleic/cytosolic protein ratios of Snail 1 were unchanged in all tested conditions. CONCLUSION: Our findings demonstrated that poly I:C accelerated collective HaCaT cell migration via autocrine/paracrine secretions of IL-8 and the subsequent incomplete epithelial-mesenchymal transition (EMT). Our findings provide a new strategy for wound healing by regulating innate immune systems in re-epithelialization.
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Interleucina-8/metabolismo , Queratinócitos/imunologia , Poli I-C/imunologia , Anticorpos Bloqueadores/metabolismo , Caderinas/metabolismo , Linhagem Celular , Movimento Celular , DNA Viral/imunologia , Transição Epitelial-Mesenquimal , Humanos , Imunidade Inata , Fatores de Transcrição da Família Snail/metabolismo , Receptor 3 Toll-Like/metabolismo , Vimentina/metabolismo , CicatrizaçãoRESUMO
Enterovirus D68 (EV-D68) is known to be causative agent of mild to severe upper and lower respiratory illnesses in sporadic cases and outbreaks. We present a case report of a 3-month-old child with acute gastroenteritis who visited a pediatric clinic in Kyushu area in Japan in 2015. A stool sample collected from the patient was screened for diarrheal viruses by multiplex RT-PCR. The result showed that the sample was positive only for enterovirus, and EV-D68 clade B3 was identified by sequence analysis of the viral protein 1 gene. This study supports an association between EV-D68 infection and acute gastroenteritis.
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Enterovirus Humano D/isolamento & purificação , Gastroenterite/microbiologia , Enterovirus , Humanos , Lactente , JapãoRESUMO
BACKGROUND: Acute encephalitis is a serious neurological condition having a high mortality rate and affecting both children and adults. This study aimed to develop a multiplex PCR method for the simultaneous screening of clinical samples for the presence of the 10 viruses presently considered as the major viral causes of acute encephalitis/ encephalopathy in Asia. METHODS: Using previously published primers that have been widely used to screen for herpes virus-6, influenza A virus, human parechovirus, herpes simplex viruses 1 and 2, Japanese encephalitis virus, group A rotavirus, enterovirus, adenovirus, and dengue virus in clinical samples, a single-tube multiplex PCR assay was developed and was tested for its sensitivity and specificity. The method was then applied to screen 57 clinical samples, consisting of 13 fecal samples, 5 throat swabs, 3 post-nasal swabs, 18 serum samples, and 18 cerebrospinal fluid (CSF) samples, collected from 18 hospitalized Japanese children with suspected viral encephalitis/encephalopathy for the target viruses, and the results were compared with those of a monoplex PCR method. RESULTS: Positive viral controls of the 10 viruses were correctly typed using this multiplex PCR method. The multiplex PCR method showed high specificity with no unspecific amplification to non-target viruses. The results of applying this PCR method for screening clinical samples showed that 6 fecal samples, 2 serum samples, and 1 CSF sample collected from 7 patients were positive for a virus, specifically group A rotavirus (4 patients, 22.2%), enterovirus (2 patients, 11.1%), or adenovirus (1 patient, 5.6%). In comparison with monoplex PCR, for group A rotavirus, enterovirus, and adenovirus, the sensitivity of this multiplex PCR method decreased for serum, cerebrospinal fluid, and throat swab samples. CONCLUSIONS: This newly developed multiplex PCR method is a simple, rapid diagnostic tool and can be used to screen clinical samples for viruses causing acute encephalitis/encephalopathy in children in Asian countries.
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DNA Viral/genética , Encefalite Viral/diagnóstico , Reação em Cadeia da Polimerase Multiplex , Vírus/genética , Doença Aguda , Calibragem , Criança , Primers do DNA , Encefalite Viral/virologia , Humanos , Japão , Reação em Cadeia da Polimerase Multiplex/normas , Valor Preditivo dos Testes , Padrões de Referência , Reprodutibilidade dos Testes , Vírus/classificaçãoRESUMO
BACKGROUND: High-mobility group box 1 (HMGB1), a DNA-binding protein, has recently been shown to have effects on HIV replication, but the effects are dependent on the cell type and the timing of infection. Using human primary T cells, this study aimed to investigate the role of HMGB1 in HIV-1 replication in newly infected cells. METHODS: Human primary T cells were infected with the HIV-1 LAI (X4) strain and then cultured in the presence of recombinant HMGB1 protein or an anti-HMGB1 antibody at various concentrations. At the indicated time points, HIV-1 p24 concentrations in the culture media were measured by ELISA. Cell proliferation, basal HMGB1 concentration, and CD3, CD4, CXCR4, and receptor for advanced glycation end products (RAGE) expression were also examined. RESULTS: Recombinant HMGB1 could enhance HIV replication in newly infected primary T cells. In the presence of an anti-HMGB1 antibody (5 µg/mL or higher), significantly lower concentrations of HIV-1 p24 were observed in the cultures of primary T cells during the post-infection period. CONCLUSIONS: The data presented suggest that HMGB1 plays a role in the enhancement of HIV-1 replication in newly infected T cells. This finding provides useful information toward understanding HIV pathogenesis and for the development of new therapeutic strategies.
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Infecções por HIV/virologia , HIV-1/crescimento & desenvolvimento , Proteína HMGB1/metabolismo , Linfócitos T/virologia , Replicação Viral , Anticorpos/farmacologia , Proliferação de Células , Células Cultivadas , Proteína do Núcleo p24 do HIV/metabolismo , Infecções por HIV/genética , Infecções por HIV/metabolismo , HIV-1/efeitos dos fármacos , HIV-1/metabolismo , Proteína HMGB1/antagonistas & inibidores , Interações Hospedeiro-Patógeno , Humanos , Cultura Primária de Células , Transdução de Sinais , Linfócitos T/efeitos dos fármacos , Linfócitos T/metabolismo , Fatores de Tempo , TransfecçãoRESUMO
BACKGROUND: Infection with Campylobacter jejuni and C. coli are recognized as the major cause of bacterial gastroenteritis in humans. METHODS: A total of 310 fecal samples collected from Thai adult patients with diarrhea in 2008 were screened for the presence of Campylobacter by PCR. Resistance to fluoroquinolone and macrolides of the detected Campylobacter strains were analyzed by studying the mutations in the gyrA and 23S rRNA genes, respectively. RESULTS: Campylobacter species were detected in 4/310 (1.3%) of diarrheal patients, and C. jejuni was found in 3 of the 4 cases (75%). Fluoroquinolone resistance was noted in 2 cases (50%); however, no resistance to macrolides was observed. CONCLUSIONS: Campylobacter was detected in a low prevalence in adult Thai patients hospitalized with diarrhea, and the resistance to fluoroquinolones is still a matter of concern in case antibiotic therapy is required.